水生生物学报2024，Vol.48Issue(3)：461-468,8.DOI:10.7541/2024.2023.0231
草鱼nrf1基因的克隆与表达及其应激响应研究
CLONING,EXPRESSION AND STRESS RESPONSE OF NRF1 GENE IN GRASS CARP(CTENOPHARYNGODON IDELLA)
摘要
Abstract
Nuclear erythroid 2 related factor 1(nrf1),a transmembrane transcription factor,plays an important role in anti-oxidative stress in animals.In this study,the nrf1 gene sequence of grass carp(Ctenopharyngodon idella)was obtained by homologous cloning,with an open reading frame(ORF)spanning 1560 bp which encoded a polypeptide consisting of 519 amino acids.Phylogenetic tree analysis showed that nrf1 was closely related to Megalobrama ambly-cephala.The tissue expression analysis of nrf1 revealed the liver as the primary site of nrf1 expression,followed by the heart and intestine.Circadian rhythm analysis showed that the expression level of nrf1 was the highest at 9:00 and significantly higher than that at 3:00,12:00,18:00,21:00 and 24:00(P＜0.05).After 24h and 48h of acute ammonia stress treatment,the expression of nrf1 gene was significantly upregulated in both low ammonia nitrogen(5 mg/L)and high ammonia nitrogen(20 mg/L)groups compared to the control group(0)(P＜0.05).The expression level of nrf1 was significantly lower in the low ammonia nitrogen group compared to the high ammonia nitrogen group at 24h(P＜0.05),and significantly higher than that in the high ammonia nitrogen group at 48h(P＜0.05).Furthermore,growth experi-ments on grass carp were conducted using three distinct protein sources(fish meal,rapeseed meal,and soybean meal),revealing that the expression level of nrf1 was significantly higher in the rapeseed meal and soybean meal groups compared to the fish meal group at 14d,28d,and 35d after feeding trial(P＜0.05).At 28d,the expression level of nrf1 in the soybean meal group was notably higher than that in the rapeseed meal group.In conclusion,the expression of the nrf1 gene in grass carp exhibits tissue specificity and is regulated by ammonia nitrogen concentration in water as well as dietary protein source.This study provides a theoretical framework for elucidating the molecular characteristics of the nrf1 gene in fish and its pivotal role in mediating antioxidative stress response.关键词
nrf1/基因克隆/氨氮胁迫/蛋白源/应激反应/草鱼Key words
nrf1/Gene cloning/Ammonia exposure/Protein source/Stress reaction/Ctenopharyngodon idella分类
生物学引用本文复制引用
莫郁坚,瞿符发,唐建洲,刘臻,曹申平,易成琳,肖扬波,杨立权,宋世航,湛瑞杰,夏浩云,孙琳,周罗璟..草鱼nrf1基因的克隆与表达及其应激响应研究[J].水生生物学报,2024,48(3):461-468,8.基金项目
国家自然科学基金(32102813) （32102813）
湖南省自然科学基金(2021JJ40627)资助[Supported by the National Natural Science Founda-tion of China(32102813) （2021JJ40627）
Hunan Provincial Natural Science Foundation of China(2021JJ40627)] （2021JJ40627）
